家畜繁殖生物技术研究现状及趋势

牛胚胎移植技术已趋于成熟。我国新疆牧科院用一步细管法移植牛冻胚受胎率达41%;牛和羊鲜胚四分胚移植也产下1头犊牛和6只羔羊。家畜体外受精,因卵子体外成熟和受精卵体外培养尚不过关,目前仍停留在实验室阶段。胚胎性别鉴定,1990年Koopman发现单拷贝基因,该基因是Y染色体的性决定区,命名为SRY,可利用PCR技术制成雄性特异DNA探针盒,进行马、牛、羊、猪早期胚胎性别鉴定。北京农学院等单位用PCR扩增牛SRY序列进行奶牛胚胎性别鉴定准确率达100%。英、日、法等国已获得牛胚胎细胞核移植后代;我国也获得1只核移植兔。北农大和新疆牧科院合作以绵羊精子为载体导入牛生长激素基因rMTbGH DNA成功,外源基因整合率为3%。     

影响绵羊冻胚移植妊娠率的因素分析

本研究对胚胎质量、胚胎移植数量、胚胎发育阶段、冷冻方法、移植方法及饲养管理水平等因素对移植产羔率的影响进行研究,以提高绵羊胚胎移植效率。结果表明:采用1.5mol/L乙二醇做冷冻保护剂对胚胎进行常规冷冻,冻前A级胚胎冷冻/解冻后可用胚率、存活率、降级率均高于B级,差异极显著(P<0.01)。移植1枚冷冻解冻后A级、B级或2枚(A+B)胚胎的受体移植产羔率分别为44.48%、46.84%和44.81%。经χ2检验分析,三者之间无显著差异(P>0.05)。移植双胚的受体双羔率为22(%53/241),以胚胎为单位计算,产羔率仅为33.40%。A级囊胚和桑椹胚的产羔率之间无显著差异(P>0.05)。1.5 mol/L乙二醇常规冷冻保存的体内胚胎移植产羔率高于EFS40玻璃化冷冻保存,但经统计学分析,二者无显著差异(P>0.05)。子宫手术移植和腹腔内窥镜法子宫移植之间产羔率不具统计学差异(P>0.05)。在牧场条件下大规模移植的平均产羔率为43.95%,范围在20%￣70%之间,受体饲养管理水平对绵羊移植产羔率有较大影响。     

碧峰峡森林土壤性状研究

碧峰峡不仅是旅游景点,还是多学科研究的天然“实验室”,地处雅安亚热带区,原始森林植被保护完好,土壤垂直分布明显,地质现象复杂,也是大熊猫、金丝猴出没之地。经研究结果：土壤呈酸性,自然肥力高．经相关分析结果：土壤有机质含量和ＣＥＣ等有关理化特性的相关性,较物理性粘粒与ＣＥＣ间更加显著。     

关联度分析在农业生态系统技术调控上的应用

农业生态系统是个“灰箱”.我们运用模糊数学的关联度分析,尝试对这类灰色系统的投入进行技术调控,并取得了良好的效果.通过对农田子系统投入的化肥(尤其是N化肥)、粪肥和还田秸秆进行技术调控的实例证明;它可以作为能、物流分析的辅助手段,互相取长补短.通过关联序分析和关联系数分析,来调整系统的投入结构和投入技艺(时、空、量级及方式的选择).本文提出了应用关联度分析的基本程序和要领以及“贡献曲线”、“潜力曲线”等概念.     

A grower-friendly method to transfer predacious mites to commercial orchards

The establishment of predacious mites in commercial orchards may be accelerated by the transfer of pruned wood in winter and summer from donor orchards to release orchards. Following winter pruning, 3-year-old and older wood is collected and transported as soon as possible in bundles to a release orchard for distribution. If the release orchard is composed of dwarf trees, then one or two bundles of 5 kg each are placed vertically at the base of the trunk of every tree in the block (0.5 to 1 ha); if the trees are of standard size, then four or five bundles used. Following summer pruning, annual shoots and suckers are distributed immediately in a release orchard composed of dwarf trees by placing 12–15 branches on the foliage of fruit-bearing branches; if the release orchard is composed of standard trees, then 50 branches are used. The pruned wood should have 20–25 leaves and not less than one predator per leaf. The release orchard should have a light infestation (two or three mites per leaf) of pest tetranychids. These phytophagous mites would serve as food and help establish the predators. The release orchard grower should develop a pest management program based on the same groups of pesticides used in the donor orchard. http://www.phytoparasitica.org posting Aug. 31, 2005.     

不同地下水位植物蒸腾耗水特性研究

在甘肃省民勤治沙站利用非称量测渗仪研究了梭梭、柠条等 10种沙区植物的蒸腾耗水特征。结果表明 ,各种植物 5a的总蒸腾量由大到小排列为 :沙枣、花棒、沙木蓼、柠条、梭梭、白皮沙拐枣 ;随植物年龄增长 ,蒸腾系数有上升的趋势 ,而蒸散系数有下降的趋势 ;在民勤沙区 6～ 9月是主要蒸腾季节 ,该时段的蒸腾量占全年蒸腾量的80 %～ 90 % ;植物的蒸腾量随年龄的增长而迅速增加 ,但增长幅度随物种不同而异。一般生长快的植物增长快 ,生长慢的植物增长慢。     

闭锁群体BLUP选择下遗传方差和近交系数的变化

采用计算机随机模拟方法模拟了在一个闭锁群体内连续对单个性状进行 1 5个世代选择的情况。选择过程中世代不重叠 ,每个世代的种畜根据动物模型最佳线性无偏预测 (BLUP)法估计的育种值进行选留 ,并在此基础上系统地比较了不同群体规模、公母比例和性状遗传力对群体遗传方差和近交系数变化的影响。结果表明 ,扩大育种群规模、增加公畜比例以及对低遗传力性状进行选择时 ,群体遗传方差降低的速度和近交系数上升的速度会更慢 ,在长期选择时可望获得更大的持续进展和适宜的近交增量     

Effect of EGFP gene transfection on the cell cycle distribution of primary cultured human chondrocytes

AIM: To investigate the effect of enhanced green fluorescence protein (EGFP) gene transfection on the cell cycle distribution of primary cultured human chondrocytes in order to establish a tracking method of cultured human nasoseptal chondrocytes. METHODS: pEGFP-N1 plasmid was amplified in E.coli, and purified by high purity kit. Primary cultured human chondrocytes,which were initially obtained from the nasoseptal cartilage, were cultured in vitro and transferred with pEGFP-N1 by means of electroporation with Amaxa nucleofector device. Transfering process and transient expression were evaluated by laser scanning confocal microscope (LSCM), the transfer efficiency and the cell cycle distribution were evaluated by flow cytometry. RESULTS: There was significant expression of EGFP at 24 h after transferring. The transfection efficiency of pEGFP-N1 into primary cultured human chondrocytes reached 35.37％ at 48 h. It didn't affect the process of cell adherance and had no effect on the cell cycle distribution. CONCLUSION: Primary cultured human chondrocytes, which were transfected with pEGFP, are alive in vitro, and the transferring process doesn't affect the cell cycle distribution. These results suggest that pEGFP-N1 is an ideal transient expression vector for primary cultured human chondrocytes and it might be a well tracer in construction tissue engineered cartilage.     

Meiotic competence of mouse oocytes reconstructed by replacing the germinal vesicle with a male pronucleus and somatic nucleus

The present study examines the contribution of the nucleus to meiotic competence in mouse oocytes that were reconstructed using nuclear transfer. Three types of reconstructed oocytes were produced: MP‐GV, by transplanting the male pronucleus (MP) into germinal vesicle (GV) stage oocytes; 3T3‐GV, by transplanting the nucleus of a National Institute of Health (NIH) 3T3 cell into a GV stage oocyte; and 3T3‐MII, by transplanting the nucleus of an NIH 3T3 cell into a metaphase II (MII) stage oocyte. The fusion rates differed, but not significantly, in the MP‐GV, 3T3‐GV, and 3T3‐MII groups (77, 63, 56%, respectively). Then, meiotic competence was compared in MP‐GV, 3T3‐GV and non‐manipulated GV stage oocytes as a control. Nuclear envelope breakdown occurred in all the reconstructed oocytes, as well as the control ones. The percentage of first polar body extrusion differed between the MP‐GV (100%), 3T3‐GV (72%), and control (67%) groups. DNA staining with Hoechst 33342 revealed that in the MP‐GV‐group oocytes that had reached MII stage, the chromosomes were condensed and aligned in a regular array similar to the normal metaphase plate. By contrast, in 3T3‐GV group oocytes, the condensed chromosomes were irregularly scattered in the cytoplasm. These results suggest that the donor nucleus affects meiotic competence in reconstructed oocytes.     

Significance of local immunity in hen reproductive organs

The current paper describes aspects of local immunity in the ovary and oviduct, and the significance of immunity to reproductive functions in hens. The immunocompetent cell populations in the ovary and oviduct change with a positive correlation to sexual activity, and gonadal steroid is one of the key factors in the increase. Local immune responses mediated by major histocompatibility complex class II and T cell subsets occur in response to infection by Salmonella enteritidis, which may contaminate eggs. In the ovary, immunocompetent cells are also suggested to play roles in the regulation of ovarian functions. Macrophages and T cells are likely to enhance the regression of atretic follicles to maintain the ovarian tissue microenvironment. Autoantibodies to ovarian tissues appeared in the hens with low egg laying frequency, suggesting that the auto‐antibodies may be one of the factors in the decline of egg production. In the oviduct, local immunity possibly has a role in the selection of sperm, though the immunoreactions may also affect sperm survival leading to the decline in fertility. The concentration of yolk IgY, which plays a role in maternal immunity transmission, significantly decreases with the aging of birds, whereas it is significantly increased by estrogen. Therefore, the immune system plays significant roles not only in defense against infection, but also in the functions of reproductive organs. Investigations on the local immune system in the reproductive organs and factors affecting it are of importance for the production of sterile eggs and improvement of reproductive functions.